背景:整合素连接激酶(ILK)通过调节Hippo-Yes相关蛋白1(YAP)途径在实体瘤中至关重要。这项研究旨在揭示幽门螺杆菌如何影响ILK水平及其在幽门螺杆菌诱导的胃癌中调节YAP的作用。
方法:构建了具有稳定的Ilk敲低和过表达的GES-1细胞和幽门螺杆菌感染的小鼠致癌模型。还有ILK,磷酸化哺乳动物STE20样蛋白激酶1(MST1),大肿瘤抑制因子1(LATS1;S909,T1079),在细胞中检测到YAP(S109,S127),和小鼠通过蛋白质印迹,免疫荧光法检测YAP的荧光强度。YAP下游基因Igfbp4和Ctgf,病理变化与肿瘤坏死因子α(TNF-α),白细胞介素-6(IL-6),白细胞介素-1β(IL-1β),实时荧光定量PCR检测小鼠胃组织一氧化氮(NO)水平,H&E,和ELISA测定。
结果:在这项研究中,稳定的Ilk敲低细胞表现出显著较高的MST1、LATS1和YAP磷酸化水平,以及GES-1细胞核中YAP的增加。相反,具有Ilk过表达的细胞显示相反的结果。幽门螺杆菌感染导致小鼠胃上皮细胞中ILK水平降低,但胃癌细胞系(MGC803,SGC7901)和胃癌组织中ILK水平升高。用ILK抑制剂OST-T315治疗会升高磷酸化MST,LATS1和YAP级别,并抑制44、48周龄小鼠Igfbp4和Ctgf的mRNA水平。OST-T315还减少了TNF-α的释放,IL-6,IL-1β,和不,以及幽门螺杆菌和N-亚硝基-N-甲基脲(NMU)治疗引起的胃癌进展。
结论:在胃肿瘤发生信号开始时,幽门螺杆菌增加ILK水平并抑制河马信号,从而促进YAP激活和胃癌进展。ILK可以作为阻止幽门螺杆菌诱导的胃癌的潜在预防靶标。
BACKGROUND: Integrin-linked kinase (ILK) is crucial in solid tumors by regulating the Hippo-Yes-associated protein 1 (YAP) pathway. This study aimed to uncover how Helicobacter pylori influences ILK levels and its role in regulating YAP during H. pylori-induced gastric cancer.
METHODS: GES-1 cells with stable Ilk knockdown and overexpression and a mouse carcinogenesis model for H. pylori infection were constructed. And ILK, the phosphorylated mammalian STE20-like protein kinase 1 (MST1), large tumor suppressor 1 (LATS1; S909, T1079), and YAP (S109, S127) were detected in cells, and mice by western blotting, as well as fluorescence intensity of YAP were assayed by immunofluorescence. YAP downstream genes Igfbp4 and Ctgf, the pathological changes and tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), interleukin-1beta (IL-1β), and nitric oxide (NO) levels in mice gastric tissues were detected by real-time PCR, H&E, and ELISA assays.
RESULTS: In this study, stable Ilk knockdown cells exhibited significantly higher phosphorylated levels of MST1, LATS1, and YAP, as well as increased YAP in the nuclei of GES-1 cells. Conversely, cells with Ilk overexpression showed opposite results. H. pylori infection led to decreased ILK levels in gastric epithelial cells but increased ILK levels in gastric cancer cell lines (MGC803, SGC7901) and gastric cancer tissues in mice. Treatment with the ILK inhibitor OST-T315 elevated the phosphorylated MST, LATS1, and YAP levels, and inhibited the mRNA levels of Igfbp4 and Ctgf at 44, 48 week-aged mice. OST-T315 also reduced the release of TNF-α, IL-6, IL-1β, and NO, as well as the progression of gastric cancer caused by H. pylori and N-Nitroso-N-methylurea (NMU) treatment.
CONCLUSIONS: Upon initiation of gastric tumorigenesis signals, H. pylori increases ILK levels and suppresses Hippo signaling, thereby promoting YAP activation and gastric cancer progression. ILK can serve as a potential prevention target to impede H. pylori-induced gastric cancer.